J Appl Biomed 14:211-219, 2016 | DOI: 10.1016/j.jab.2016.02.001

Caspase dependent apoptotic activity of Rubus fairholmianus Gard. on MCF-7 human breast cancer cell lines

Blassan P. Georgea,*, Heidi Abrahamsea, Thangaraj Parimelazhaganb
a Laser Research Centre, Faculty of Health Sciences, University of Johannesburg, P.O. Box 17011, Doornfontein 2028, South Africa
b Bioprospecting Laboratory, Department of Botany, Bharathiar University, Coimbatore 641 046, Tamil Nadu, India

Developing plant derived chemopreventive agents that can divert the carcinogenic process and inhibit tumour progression may greatly reduce serious health consequences of cancer. This study investigated the antiproliferative and apoptosis inducing ability of Rubus fairholmianus root acetone (RFRA) on MCF-7 cells. RFRA treatments showed an increase in the in vivo antioxidant levels such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione (GSH) in mice. The extract showed significant antitumour activity with 76.57% increase in life span of ascites tumour bearing mice and 1.94 cm3 reductions in solid tumour volume at 250 mg/kg. The cytotoxicity spectrum analysis indicated that RFRA is a potent cytocidal agent with varying levels of toxicity (IC50: 57.14-29.36 μg/ml). A significant dose dependent decrease in viability, proliferation and increase in cytotoxicity, caspase 3/7 activity were observed in MCF-7 cells. RFRA (20 μg/ml) induced apoptosis in MCF-7 cells, which was observed by morphology; DNA ladder formation and increased apoptotic population in flow cytometry analysis. These findings strongly suggest the use of R. fairholmianus as natural antioxidant with profound antitumour activities. It can act as a potent cytotoxic agent, which is able to induce apoptosis in MCF-7 cells via caspase 3/7 upregulation.

Keywords: Rubus fairholmianus; Antioxidant; Antitumour; Cytotoxicity; Annexin V-FITC, PI; Caspase; Apoptosis

Received: October 9, 2015; Revised: December 1, 2015; Accepted: February 8, 2016; Published: August 1, 2016  Show citation

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George BP, Abrahamse H, Parimelazhagan T. Caspase dependent apoptotic activity of Rubus fairholmianus Gard. on MCF-7 human breast cancer cell lines. J Appl Biomed. 2016;14(3):211-219. doi: 10.1016/j.jab.2016.02.001.
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References

  1. Aebi, M., 1974. Catalase estimation. In: Berg Meyer, H.V. (Ed.), Methods of Enzymatic Analysis. Verlag Chemie, New York, pp. 673-684. Go to original source...
  2. Bagchi, D., Swaroop, A., Preuss, H.G., Bagchi, M., 2014. Free radical scavenging, antioxidant and cancer chemoprevention by grape seed proanthocyanidin: an overview. Mutat. Res. 768, 69-73. Go to original source... Go to PubMed...
  3. Balkwill, F.R., Mantovani, A., 2012. Cancer-related inflammation: common themes and therapeutic opportunities. Semin. Cancer Biol. 22, 33-40. Go to original source... Go to PubMed...
  4. Barukial, J., Sarmah, J.N., 2011. Ethnomedicinal plants used by the people of Golaghat district, Assam, India. Int. J. Med. Aromat. Plants 1, 203-211.
  5. Bhandari, P.R., 2015. Crocus sativus L. (saffron) for cancer chemoprevention: a mini review. J. Tradit. Complement. Med. 5, 81-87. Go to original source... Go to PubMed...
  6. Campling, B.G., Pyn, J., Baker, H.M., Lan, Y.M., 1991. Chemosensitivity testing of small cell lung cancer using the MTT assay. Br. J. Cancer 63, 75-83. Go to original source... Go to PubMed...
  7. Earnshaw, W.C., Martins, L.M., Kaufmann, S.H., 1999. Mammalian caspases: structure, activation, substrates, and functions during apoptosis. Annu. Rev. Biochem. 68, 383-424. Go to original source... Go to PubMed...
  8. Elmore, S., 2007. Apoptosis: a review of programmed cell death. Toxicol. Pathol. 35, 495-516. Go to original source... Go to PubMed...
  9. Finn, C.E., 2008. Rubus spp., blackberry. In: Janick, J., Paull, R.E. (Eds.), The Encyclopaedia of Fruits and Nuts. CABI, Cambridge, MA, pp. 348-351.
  10. Firdous, A.P., Preethi, K.C., Kuttan, R., 2010. Antioxidant potential of meso-zeaxanthin a semi synthetic carotenoids. Food Chem. 119, 1096-1101. Go to original source...
  11. George, B.P., Parimelazhagan, T., Chandran, R., 2013a. Evaluation of total phenolic content, antioxidant and analgesic potential of Rubus fairholmianus Gard. IJPPS 5, 484-488.
  12. George, B.P., Parimelazhagan, T., Chandran, R., 2014a. Antiinflammatory and wound healing properties of Rubus fairholmianus Gard. root - an in vivo study. Ind. Crops Prod. 54, 216-225. Go to original source...
  13. George, B.P., Parimelazhagan, T., Yamini, T.K., Sajeesh, T., 2013b. Antitumour and wound healing properties of Rubus ellipticus Smith. J. Acupunct. Meridian Stud., http://dx.doi.org/10.1016/j.jams.2013.10.002. Go to original source... Go to PubMed...
  14. George, B.P., Thangaraj, P., Sulaiman, C., Piramanayagam, S., Ramaswamy, S.K., 2014b. Bioassay directed isolation and biological evaluation of compounds isolated from Rubus fairholmianus Gard. Biomed. Res. Int., http://dx.doi.org/10.1155/2014/204340 Article ID 204340. Go to original source... Go to PubMed...
  15. George, B.P., Tynga, I.M., Abrahamse, H., 2015. In vitro antiproliferative effect of the acetone extract of Rubus fairholmianus Gard. root on human colorectal cancer cells. Biomed. Res. Int., http://dx.doi.org/10.1155/2015/165037 Article ID 165037. Go to original source... Go to PubMed...
  16. Grankvist, K., Lernmark, A., Taljedal, I.B., 1977. Alloxan cytotoxicity in vitro. Microscope photometric analyses of Trypan Blue uptake by pancreatic islet cells in suspension. Biochem. J. 162, 19-24. Go to original source... Go to PubMed...
  17. Hafeman, D.G., Sundae, R.A., Houestra, W.G., 1974. Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. J. Nutr. 104, 580-587. Go to original source... Go to PubMed...
  18. Hansakul, P., Aree, K., Tanuchit, S., Itharat, A., 2014. Growth arrest and apoptosis via caspase activation of dioscoreanone in human non-small-cell lung cancer A549 cells. BMC Complement. Altern. Med. 14, 413, http://dx.doi.org/10.1186/1472-6882-14-413. Go to original source... Go to PubMed...
  19. Herrmann, H., Lorenz, H.M., Voll, R., 1994. A rapid and simple method for the isolation of apoptotic DNA fragments. Nucleic Acids Res. 22, 5506-5507. Go to original source... Go to PubMed...
  20. Jeong, J.H., Jung, H., Lee, S.R., Lee, H.J., Hwang, K.T., Kim, T.Y., 2010. Anti-oxidant, anti-proliferative and anti-inflammatory activities of the extracts from black raspberry fruits and wine. Food Chem. 123, 338-344. Go to original source...
  21. Kim, E.J., Lee, Y.J., Shin, H.K., Park, J.H., 2005. Induction of apoptosis by the aqueous extract of Rubus coreanum in HT-29 human colon cancer cells. Nutrition 21, 1141-1148. Go to original source... Go to PubMed...
  22. McCord, J.M., Fridovich, I., 1969. Superoxide dismutase enzyme function for erythrocuprein. J. Biochem. 244, 6049-6056. Go to original source...
  23. Monga, J., Pandit, S., Chauhan, C.S., Sharma, M., 2013. Cytotoxicity and apoptosis induction in human breast adenocarcinoma MCF-7cells by (+)-cyanidan-3-ol. Exp. Toxicol. Pathol. 65, 1091-1100. Go to original source... Go to PubMed...
  24. Moron, M.A., Depierre, J.W., Vick, M.B., 1979. Levels of glutathione, glutathione reductase and glutathione-Stransferase activities in rat liver. Biochim. Biophys. Acta 582, 67-68. Go to original source... Go to PubMed...
  25. Naselli, F., Tesoriere, L., Caradonna, F., Bellavia, D., Attanzio, A., Gentile, C., Livrea, M.A., 2014. Anti-proliferative and proapoptotic activity of whole extract and isolated indicaxanthin from Opuntia ficusindica associated with reactivation of the onco-suppressor p16INK4a gene in human colorectal carcinoma (Caco-2) cells. Biochem. Biophys. Res. Commun. 450, 652-658. Go to original source... Go to PubMed...
  26. Organization for Economic Co-operation and Development, 2001. OECD Guidelines for the Testing of Chemicals, Revised Draft Guidelines, vol. 423. OECD, USA.
  27. Rajeshkumar, N.V., Joy, K.L., Kuttan, G., Ramsewak, R.S., Nair, M.G., Kuttan, R., 2002. Antitumour and anticarcinogenic activity of Phyllanthus amarus extract. J. Ethnopharmacol. 81, 17-22. Go to original source... Go to PubMed...
  28. Ruby, A.J., Kuttan, G., Babu, D.K., Rajasekharan, K.N., Kuttan, R., 1995. Anti-tumour and antioxidant activity of natural curcuminoids. Cancer Lett. 94, 79-83. Go to original source... Go to PubMed...
  29. Saad, S., Dakwar, O., Said, G., Abu-Hijleh, G., Battah, F.A., Kmeel, A., Aziazeh, H., 2006. Evaluation of medicinal plant hepatotoxicity in co-cultures of hepatocytes and monocytes. J. Evid. Based Complement. Altern. Med. 3, 93-98. Go to original source... Go to PubMed...
  30. Seeram, N.P., Adams, L.S., Zhang, Y.J., Lee, R., Sand, D., Scheuller, H.S., Heber, D., 2006. Blackberry, black raspberry, blueberry, cranberry, red raspberry, and strawberry extracts inhibit growth and stimulated apoptosis of human cancer cells in vitro. J. Agric. Food Chem. 54, 9329-9339. Go to original source... Go to PubMed...
  31. Sulaiman, C.T., Gopalakrishnan, V.K., Balachandran, I., 2014. Chromatographic and spectrophotometric studies in selected Acacia species. J. Planar Chromatogr. 27, 88-92. Go to original source...
  32. Vidau, C., Brunet, J.L., Badiou, A., Belzunces, L.P., 2009. Phenylpyrazole insecticides induce cytotoxicity by altering mechanisms involved in cellular energy supply in the human epithelial cell model Caco-2. Toxicol. In Vitro 23, 589-597. Go to original source... Go to PubMed...